We propose to acquire an Asylum Research MFP-3D atomic force microscope (AFM), which has high- resolution force spectroscopy and scanning capabilities, integrated into a Nikon TI-E epifluorescence microscope for a combined AFM/microscope instrument. This MFP-3D/Nikon instrument can apply calibrated forces ranging between pico- and nano-newton, a force range relevant to many biological processes. Unlike our current stand-alone non-imaging MFP-1D AFM instrument, the combined MFP-3D/Nikon instrument allows for both imaging and non-imaging applications. The MFP-3D/Nikon instrument will directly support a wide range of applications that have recently been funded by the NIH. These applications include (1) Mechanistic studies of the defects in cell mechanics induced by disease-causing mutations in the Lmna gene by Dr. Wirtz's group (1R01GM084204), (2) Biophysical studies of selectin-mediated cell-cell adhesion in cancer metastasis by Dr. Konstantopoulos' group (5R01CA101135), (3) High-resolution imaging of surfaces covered with engineered bacterial cells by Dr. Ostermeier's group 5R01GM066972), (4) Biophysical studies of the mechanical role of cytoskeletal proteins in controlling bacterial cell shape and mediating cell division by Dr. Sun's group (5R01GM075305), as well as (5) Single-molecule force spectroscopy studies of cadherin- mediated cell-cell adhesion by Dr. Wirtz group (1R21CA13789). This combined instrument will be placed in an 800 sq ft state-of-the-art facility NanoBio Lab of the Institute for NanoBioTechnology (INBT) in the NEB Building at the Johns Hopkins University Homewood campus. This core facility has a demonstrated outstanding record of providing excellent imaging service to the INBT research community, offering highly popular laboratory courses in nanoscale science and engineering for applications in biology and medicine, and participating in large-scale outreach efforts. Housing the combined AFM/microscope in the INBT NanoBio Lab directed by D. Wirtz will greatly enhance its efficient and balanced use by the four primary NIH- funded users and other users under the supervision of an advisory committee. The INBT will provide support for the maintenance of the instrument, will offer the laboratory space, and will provide the administrative/accounting/web support for the management of the facility, scheduling of users, and financial reporting. PUBLIC HEALTH RELEVANCE: The requested MFP-3D/Nikon instrument will be used in a wide range of biomedical science projects of importance to human diseases, including cancer, laminopathies, and bacterial and viral infection.